Long Term Goals
miRNA Profile
- Read literature
- Choose profile
Building the Cell Detector Circuit
- Build Choose profile
- Verify sensors
- Create mathematical model
- Make test bed
- Controlled miRNA expression
- Constructed miRNA sensors
Mini experiments
- Single miRNA detector circuit
Experiments
- L7ae repression
- TAL14 repression
- TAL21 repression
- Single input sensors
- Multiple input sensors
Alzheimer's Model
- Agree on a team wide, full Alzheimer's model
Today's Agenda
How To Please Ron/Brain
- Keep updating the parts tree (include experiments)
- Meet w/ Kyle
- FlowJo
- Bright Field Images
miRNA Sensors (Theory)
- Troubleshoot
- What's going on with the double input low sensor
- Is there a different GG thermal cycler program for Bbs1?
- Run new transfection plate/protocol by Brian
- Move siRNA into our cryobox
- Grab sequencing primer from Jeremy's bench
- Re-analyze all of our data in MATLAB (Gary)
- Learn how to acquire MATLAB from Brian
- Find natural concentrations of relevant miRNAs.
miRNA Sensors (Lab Work)
- Pick colonies
- pEXPR: MAV1212-hEF1a-TAL14 (Mini)
- Give plates to Brian
- pENTR: TAL14p
- pENTR: TAL21p
- pENTR: Something for yeast for Brian
- Restriction Digest & run gel
- pEXPR: MAV1212-TRE-L7ae (LR)
- pEXPR: MAV1212-TRE-GAL4VP16 (LR)
- Transform
- pEXPR: MAV1212-hEF1a-GAL4VP16
- Golden Gate
- All the sensors
- Split and Seed 4 plates
NOTES
- Midi-prep hEF1a:rtTA and hEF1a: Gal4VP16 2A rtTA3
- Mini-prep MAV1212 hEF1A Tal14
- Digest and Map MAV1212 hEF1A Tal14
- Transform third LR trial if not verified
- Pick/Streak hEF1a: Gal4VP16 2A rtTA3
NOTES:
hEF1a: Gal4VP16 2A rtTA3 culture did not grow so We transfected way too much DNA --> new protocol with 1 uL total